My MSc project aims to develop Nicotiana benthamiana (a tobacco relative) as an industrial-scale production system for the anti-diabetic compound Montbretin A (MbA). MbA production was previously achieved in N. benthamiana by Agrobacterium-mediated infiltration of montbretia genes. However, it is necessary to greatly increase MbA yields to meet anticipated demand. To this end, I applied metabolomic, transcriptomic, bioinformatic, and in vitro and in planta enzyme assay approaches to study the metabolism of MbA-bioengineered N. benthamiana plants. I used this information to inform two strategies for improving MbA yield. (1) When expressed in N. benthamiana, the MbA pathway has numerous bottlenecks that limit MbA yield. By increasing the expression of the enzymes that come after these bottlenecks, I decreased intermediate accumulation and improved MbA yields. (2) MbA-bioengineered N. benthamiana also accumulates numerous undersired side-products. I identified the enzymes responsible for some of these sideproducts and cloned RNAi silencing constructs with the goal of reducing side-product formation and redirecting flux towards MbA. While working on my own research, I also mentored other students, providing training for experimental design, wet lab techniques, instrument operation, and data analysis.

I visited the Irmisch lab as part of my MSc research in the Bohlmann lab. During my time in Leiden, I primarily focused on addressing the question of side-product formation in MbA-bioengineered N. benthamiana. To this end, I applied bioinformatic, metabolomic, and in vitro assay approaches to identify undesired side-products and to identify and characterize some of the UDP-glycosyltransferases responsible for their formation.

iGEM is a worldwide engineering biology competition for student-led teams that apply molecular biology, engineering approaches, and social context to important world issues.Following 3 years of competing in iGEM as a student, I returned to serve as a judge for the 2021 and 2022 iGEM competitions.I thoroughly reviewed teams’ documentation and presentations, applying previous knowledge of synthetic biology and experience as an iGEM participant to provide constructive critique and feedback. I also served as the chair of several judging sessions.

Used metabolic engineering strategies to develop E. coli and Corynebacterium glutamicum as production systems for sustainable textile dyesUsed Gibson assembly to build an E. coli-C. glutamicum shuttle vector encoding dye biosynthesisTroubleshot C. glutamicum transformation using various heat shock and electroporation protocols Optimized fermentation conditions for dye production by varying the length of the fermentation and supplementing growth media with upstream metabolites

Used metabolic engineering to increase production of industrially-relevant isoprenoids in E. coliEmployed CRISPR tools to insert variations of a synthetic isopentenol-utilisation pathway into the E. coli chromosome for reduced metabolic burden and stable maintenanceDesigned CRISPR insertion experiments using online tools and used Gibson assembly to build the required constructs, verifying with colony PCR and sequencingTransformed engineered strains with various carotenoid-producing plasmids and characterised the resulting carotenoids using UV/Vis spectroscopy and HPLC Mentored several other students, providing training for experimental design, wet lab techniques, instrument operation, and data analysis

iGEM is a worldwide engineering biology competition for student-led teams that apply molecular biology, engineering approaches, and social context to important world issuesThe 2020 project seeks to use metal-binding proteins to recover valuable metals from industrial wastes: Proteins will be identified, engineered to improve specificity and affinity, and immobilised in a packed column reactorWorked closely with other team leads to select a project, coordinate research activities across the 40+ member team, and navigate a challenging funding situationInterviewed, hired, trained, and supervised 14 undergraduate students in varied programs and years, focusing on engagement, retention, well-being, and community Supervised various research activities by providing guidance and direction, ensuring that work remained relevant and met deadlines Research included process design and modelling using Fenics and COMSOL and protein engineering using PyMOL, Osprey, Autodock, and in silico alanine scanningCoordinated communication and collaboration with students and faculty of many backgrounds at Waterloo and across North America and the UK, assisting less experienced teams with modelling

iGEM is a worldwide engineering biology competition for student-led teams that apply molecular biology, engineering approaches, and social context to important world issuesTeam earned a silver medal for 2019 project, which sought to confer linuron resistance to the root-nodule forming bacterium B. diazoefficiens by expressing a two-step linuron degradation pathwayGuided project direction and independently designed, prioritised, and executed experiments to test relevant hypotheses, troubleshooting extensivelyExperiments included: bacterial MIC toxicity assays, DNA construct design and assembly, biparental conjugation into B. diazoefficiens, semi-quantitative RT-PCR, flow cytometry, nodulation assays with soybean plants, and statistical and numerical data analysis

Used bioinformatic approaches to investigate the prevalence and identity of reductive dehalogenases in enrichment culture metagenomes, focusing on hexachlorocyclohexane (HCH) dechlorinating enzymesQuality control, assembly, and annotation of metagenome sequencing data were performed at the command line using MetaErg and with the online workflow manager GalaxyHCH-dechlorinating enzymes were identified and characterised by homology modelling and comparison with literature, revealing trends in enzyme distribution across enrichment cultures and their bins Audited two graduate courses: Fundamentals of Genomic Data Science and Introduction to R, using acquired skills to process and present data

Tasked with studying microbial selenate reduction using molecular biology and biochemistry approaches in an engineering-focused research group Independently developed and validated methods for extracting genomic DNA from difficult environmental and industrial samples including biofilmsResponsible for maintenance and sampling of a delicate anaerobic selenate-reducing chemostat: performed ATP measurements, COD measurements, ICP-OES, DNA extraction, and qPCR Performed extensive literature review of selenate reductases, focusing on structure, function, and phylogeny. Used bioinformatic approaches to identify possible selenate reductases from online databases, group them by phylogeny, and design degenerate qPCR primers

Involved in development of fluorescent lateral flow immunoassays for quantification of allergy-related antibodies in patient blood samples, managing rapidly changing start-up research prioritiesIndependently optimised preparation and analysis of antibody-microparticle conjugates and biotinylated allergen components for use in assay research and manufactureDeveloped and improved methods for blood filtration, manufacturing various 3D-printed filter assemblies with different types of filter pads and evaluating their effects on assay LOQ and SNR Performed numerical and statistical analyses of experimental data using Microsoft Excel and other specialised software packages, using the results to design subsequent experimentsSolved multidisciplinary problems through effective communication with assay scientists, software and hardware engineers, and business development specialists

Nominated by employer for university-wide “Co-op Student of the Year” awardIndependently designed and performed experiments based on existing literature and changing start-up research priorities with minimal supervisionCompleted multi-step organic reactions to produce polysaccharide nanomaterial therapeuticsCharacterised resulting nanomaterials by titration, NMR (1H, 13C, HMBC), IR spectroscopy, dynamic light scattering; troubleshot analytical methods to improve precision and accuracyImproved oral, written, and visual communication skills by preparing numerous technical reports, communicating effectively with management, and delivering formal oral presentations

Investigated ion exchange chromatography for the downstream purification of monoclonal antibodies, optimising process conditions to improve yield and reduce aggregationIndependently designed and performed experiments to evaluate the use of thioflavin T as an extrinsic fluorophore for quantifying and characterising antibody aggregates Employed analytical methods including absorbance and fluorescence spectroscopies, performed statistical and numerical analyses of data, and applied existing literature to interpret findings

Demonstrated resourcefulness by following protocols for syntheses of long-chain dicarboxylic acids as a high school student with little assistance or previous knowledge Improved understanding of organic synthesis techniques, laboratory safety, and analytical methods including 1H NMR spectroscopyPresented work at regional science fair and won several awards, demonstrating oral presentation skills and visual communication